Neurochemical Research

, Volume 32, Issue 3, pp 517–524

Fulminant Hepatic Failure in Rats Induces Oxidative Stress Differentially in Cerebral Cortex, Cerebellum and Pons Medulla

  • K. V. Sathyasaikumar
  • I. Swapna
  • P. V. B. Reddy
  • Ch. R. K. Murthy
  • A. Dutta Gupta
  • B. Senthilkumaran
  • P. Reddanna
Original Paper


Hepatic Encephalopathy (HE) is one of the most common complications of acute liver diseases and is known to have profound influence on the brain. Most of the studies, available from the literature are pertaining to whole brain homogenates or mitochondria. Since brain is highly heterogeneous with functions localized in specific areas, the present study was aimed to assess the oxidative stress in different regions of brain-cerebral cortex, cerebellum and pons medulla during acute HE. Acute liver failure was induced in 3-month old adult male Wistar rats by intraperitoneal injection of thioacetamide (300 mg/kg body weight for two days), a well known hepatotoxin. Oxidative stress conditions were assessed by free radical production, lipid peroxidation, nitric oxide levels, GSH/GSSG ratio and antioxidant enzyme machinery in three distinct structures of rat brain-cerebral cortex, cerebellum and pons medulla. Results of the present study indicate a significant increase in malondialdehyde (MDA) levels, reactive oxygen species (ROS), total nitric oxide levels [(NO) estimated by measuring (nitrites + nitrates)] and a decrease in GSH/GSSG ratio in all the regions of brain. There was also a marked decrease in the activity of the antioxidant enzymes-glutathione peroxidase, glutathione reductase and catalase while the super oxide dismutase activity (SOD) increased. However, the present study also revealed that pons medulla and cerebral cortex were more susceptible to oxidative stress than cerebellum. The increased vulnerability to oxidative stress in pons medulla could be due to the increased NO levels and increased activity of SOD and decreased glutathione peroxidase and glutathione reductase activities. In summary, the present study revealed that oxidative stress prevails in different cerebral regions analyzed during thioacetamide-induced acute liver failure with more pronounced effects on pons medulla and cerebral cortex.


Oxidative stress Thioacetamide Cerebral cortex Cerebellum Pons medulla Fulminant hepatic failure 


  1. 1.
    Dawson TM, Synder SH (1994) Gases as biological messengers: nitric oxide and carbon monoxide in the brain. J Neurosci 14:5147–5159PubMedGoogle Scholar
  2. 2.
    Robberecht W (2000) Oxidative stress in amyotrophic lateral sclerosis. J Neurol Suppl 1:I1–6Google Scholar
  3. 3.
    Richardson SJ (1993) Free radicals in the genesis of Alzheimer’s disease. Ann NY Acad Sci 695:73–76PubMedCrossRefGoogle Scholar
  4. 4.
    Ben-Shachar D, Riederer P (1991) Iron-melanin interaction and lipid peroxidation: implication for Parkinson’s disease. J Neurochem 57:1609–1614PubMedCrossRefGoogle Scholar
  5. 5.
    Braughler JM, Hall ED (1989) Central nervous system trauma and stroke. Biochemical considerations for oxygen radicals formation and lipid peroxidation. J Free Radical Biol Med 6:289–301CrossRefGoogle Scholar
  6. 6.
    Ames BN, Shigenaga MK, Hagen TM (1993) Oxidants, antioxidants and the degenerative diseases of aging. Proc Natl Acad Sci 90:7915–7922PubMedCrossRefGoogle Scholar
  7. 7.
    Reiter RJ (1995) Oxidative processes and antioxidant defense mechanisms in the ageing brain. FASEB J 9:526–533PubMedGoogle Scholar
  8. 8.
    Manoli L, Gamaro GD, Silveira PP, Dalmaz C (2000) Effect of chronic variate stress on TBARS and on total radical trapping potential in distinct regions of brain. Neurochem Res 25:915–921PubMedCrossRefGoogle Scholar
  9. 9.
    Smythies J (1999) The neurotoxicity of glutamate, dopamine, iron and reactive oxygen species: functional interrelationship in health and diseases. Neurotox Res 1:27–29PubMedCrossRefGoogle Scholar
  10. 10.
    Choi JH, Yu BP (1995) Brain synaptosomal ageing: free radicals and membrane fluidity. Free Radical BiolMed 18:133–139CrossRefGoogle Scholar
  11. 11.
    Halliwell B, Gutteridge JMC (1985) Oxygen radicals and the nervous system. Trends Neurosci 8:22–26CrossRefGoogle Scholar
  12. 12.
    Halliwell B (1992) Reactive oxygen species and the central nervous system. J Neurochem 59:1609–1623PubMedCrossRefGoogle Scholar
  13. 13.
    Reznick AZ, Packer AL (1993) Free radicals and antioxidants in muscular neurological disease and disorders. In: Poli G, Albano E, Dianzani MU (eds) Free radicals: from basic science to medicine. Birkhauser Verlag, Basel, pp 425–437Google Scholar
  14. 14.
    Hill JM, Switzer RC (1984) The regional distribution and cellular localization of iron in the rat brain. Neuroscience 11:595–603PubMedCrossRefGoogle Scholar
  15. 15.
    Riordan MS, Williams R (2000) Fulminant hepatic failure: a century of Progress. Clin Liver Dis 4:25–45PubMedCrossRefGoogle Scholar
  16. 16.
    Blei AT (2005) The pathophysiology of brain edema in acute liver failure. Neurochem Int 47:71–77PubMedCrossRefGoogle Scholar
  17. 17.
    Jalan R, Shawcross D, Davies N (2003) The molecular pathogenesis of hepatic encephalopathy. Int J Biochem Cell Biol 35:1175–1181PubMedCrossRefGoogle Scholar
  18. 18.
    Norenberg DM (2003) Oxidative stress and Nitrosative stress in ammonia neurotoxicity. Hepatology 37(2):244–247CrossRefGoogle Scholar
  19. 19.
    Reddy PVB, Murthy Ch RK, Reddanna P (2004) Fulminant hepatic failure induced oxidative stress in non-synaptic mitochondria of cerebral cortex in rats. Neurosci Lett 368:15–20PubMedCrossRefGoogle Scholar
  20. 20.
    Kosenko E, Kaminsky Y, Lopata O, Muravyov N, Kaminsky A, Hermenegildo C, Felipo V (1998) Nitroarginine, an inhibitor of nitric oxide synthase, prevents changes in super oxide radical and antioxidant enzymes induced by ammonia intoxication. Metabol Brain Dis 13:29–41CrossRefGoogle Scholar
  21. 21.
    Swapna I, SathyaSaiKumar KV, Reddy PVB, Reddanna P, Murthy ChRK, Senthilkumaran B (2006) Phospholipids and cholesterol alterations accompany structural disarray in myelin membrane of rats with hepatic encephalopathy induced by thioacetamide. Neurochem Int 49:238–244PubMedCrossRefGoogle Scholar
  22. 22.
    Norton NS, McConnell JR, Rodriguez-Sierra JF (1997) Behavioral and physiological sex differences observed in an animal model of fulminant hepatic failure. Physiol Behav 62: 1113–1124PubMedCrossRefGoogle Scholar
  23. 23.
    Rahman TM, Hodgson HJF (2003) The effects of early and late administration of inducible nitric oxide synthatase in a thioacetamide induced model of acute hepatic failure in the rat. J Hepatol 38:583–590PubMedCrossRefGoogle Scholar
  24. 24.
    Gellar D (1998) A rat model of hepatic encephalopathy due to fulminant hepatic failure: the role of supportive therapy. In: Soeters PB, Wilson JHP, Meijer AJ, Holm E (eds) Advances in ammonia metabolism and hepatic encephalopathy. Elsevier Science Publishers, Amsterdam, pp 213–217Google Scholar
  25. 25.
    Muriel P, Alba N, Perez-Alvarez VM, Shibayama M, Tsutsumi VK (2001) Kupffer cells inhibition prevents hepatic lipid peroxidation and damage induced by carbon tetrachloride. Comp Biochem Physiol Part C 130:219–226CrossRefGoogle Scholar
  26. 26.
    Montoliu C, Valles S, Renau-Piqueras J, Guerri C (1994) Ethanol-induced oxygen radical formation and lipid peroxidation in rat brain: effect of chronic alcohol consumption. J Neurochem 63:1855–1862PubMedCrossRefGoogle Scholar
  27. 27.
    Sastry KVH, Moudgal RP, Mohan J, Tyagi TS, Rao GS (2002) Spectrophotometric determination of serum nitrites and nitrates by copper-cadmium alloy. Anal Biochem 306:79–82PubMedCrossRefGoogle Scholar
  28. 28.
    Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  29. 29.
    Paglia DL, Valentine WN (1967) Studies on qualitative and quantitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70:158–169PubMedGoogle Scholar
  30. 30.
    Claiborne A (1985) Catalase. In: Greenwald R. (eds) Handbook of methods for oxygen radical research. CRC Press, New York, pp 283–284Google Scholar
  31. 31.
    Carlberg I, Mannervick B (1975) Purification and characterization of the flavor enzyme glutathione reductase from rat liver. J Biol Chem 250:5474–5480Google Scholar
  32. 32.
    Bay B-H, Lee Y-K, Tan BK-H, Ling E-A (1999) Lipid peroxidative stress and anti-oxidative enzymes in brains of milk-supplemented rats. Neurosci Lett 277:127–130PubMedCrossRefGoogle Scholar
  33. 33.
    Hissin PJ, Hilf R (1976) A fluorometric method for determination of oxidized and reduced glutathione in tissues. Anal Biochem 74:214–226PubMedCrossRefGoogle Scholar
  34. 34.
    Kosenko E, Venediktova N, Kaminsky Y, Montoliu C, Felipo V (2003) Sources of oxygen radicals in brain in acute ammonia intoxication in vivo. Brain Res 981:193–200PubMedCrossRefGoogle Scholar
  35. 35.
    Kikugawa K, Oikawa N, Miyazawa A, Shindo K, Kato T (2005) Interaction of nitric oxide with glutathione or cysteine generates reactive oxygen species causing DNA single strand breaks. Biol Pharm Bull 28:998–1003PubMedCrossRefGoogle Scholar
  36. 36.
    Schliess F, Gorg B, Fischer R, Desjardins P, Bidmon HJ, Hermann A, Butterworth RF, Zilles K, Haussinger D (2002) Ammonia induces MK-801 sensitive nitration and phosphorylation of protein tyrosine residues in rat astrocytes. FASEB J 16:739–741PubMedGoogle Scholar
  37. 37.
    Hilgier W, Anderzhanova E, Oja SS, Saransaari P, Albrecht J (2003) Taurine reduces ammonia and N-methyl-d-aspartate-induced accumulation of cyclic GMP and hydroxyl radicals in microdialysate of the rat striatum. Eur J Pharmacol 468:21–25PubMedCrossRefGoogle Scholar
  38. 38.
    Wang JH, Redmond HP, Wu QD, Bouchier HD (1998) Nitric oxide mediates hepatocytes injury. Am J Physiol 275:1117–1126Google Scholar
  39. 39.
    Malcolm C, Grieve A, Ritchie L, Schousboe A, Griffiths R (1996) NMDA receptor-mediated cGMP synthesis in primary cultures of mouse cerebellar granule cells appears to involve neuron-astrocyte communication with NO operating as the intracellular messenger. J Neurosci Res 45:129–142PubMedCrossRefGoogle Scholar
  40. 40.
    Murthy ChRK, Bender AS, Dombro RS, Bai G, Norenberg MD (2000) Elevation of glutathione levels by ammonium ions in primary cultures of rat astrocytes. Neurochem Int 37:255–268CrossRefGoogle Scholar
  41. 41.
    Lu SC, Huang ZZ, Yang H, Tsukamoto H (1999) Effect of thioacetamide on the hepatic expression of gamma-glutamylcysteine synthatase subunits in the rat. Toxicol Appl Pharmacol 159:161–168PubMedCrossRefGoogle Scholar
  42. 42.
    Arivazhagan P, Shila S, Kumaran S, Panneerselvam C (2002) Effect of DL-alpha lipoic acid on the status of lipid peroxidation and antioxidant enzymes in various brain regions of rats. Exp Gerontol 37:803–811PubMedCrossRefGoogle Scholar
  43. 43.
    Dogru-Abbasoglu S, Kanbagli O, Balkan J, Cevikbas U, Aykac-Toker G, Uysal M (2001) The protective effect of taurine against thioacetamide hepatotoxicity of rats. Hum Exp Toxicol 20:23–27PubMedCrossRefGoogle Scholar
  44. 44.
    Mandavilli BS, Rao KS (1996) Neurons in the cerebral cortex are more susceptible to DNA-damage in ageing rat brain. Biochem Mol Biol Int 40:507–514PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • K. V. Sathyasaikumar
    • 1
  • I. Swapna
    • 1
  • P. V. B. Reddy
    • 1
  • Ch. R. K. Murthy
    • 1
  • A. Dutta Gupta
    • 1
  • B. Senthilkumaran
    • 1
  • P. Reddanna
    • 1
  1. 1.Department of Animal Sciences, School of Life SciencesUniversity of HyderabadHyderabadIndia

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